Anti-Cyclophilin F antibody [E11AE12BD4]

7 product images

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  Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  Lane 1: Wild type HAP1 whole cell lysate (20 μg)
  Lane 2: Cyclophilin F knockout HAP1 whole cell lysate (20 μg)
  Lane 3: HeLa whole cell lysate (20 μg)
  Lane 4: Hek293 whole cell lysate (20 μg)
  
  Lanes 1 - 4: Merged signal (red and green). Green - ab110324 observed at 24 kDa. Red - loading control, Anti-GAPDH antibody [EPR16891] - Loading Control ab181602, observed at 37 kDa.
  

  ab110324 detected the expected band for Cyclophilin F in wild type HAP1 cells and the band was not seen in Cyclophilin F knockout HAP1 cells. Wild-type and Cyclophilin F knockout samples were subjected to SDS-PAGE. ab110324 and Anti-GAPDH antibody [EPR16891] - Loading Control ab181602 (Rabbit anti GAPDH loading control) were incubated overnight at 4°C at 1 ug/ml and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye^® 800CW) preabsorbed Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye^® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  Predicted band size: 22 kDa

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  Immunocytochemistry/ Immunofluorescence - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  Immunocytochemistry analysis using ab110324 at 1μg/ml staining Cyclophilin 40 in Cultured Human embryonic lung-derived fibroblasts (strain MRC5), (fixed, treated for heat-induced antigen retrieval, permeabilized) followed by an AlexaFluor® 488-conjugated-goat-anti-mouse IgG1 isotype specific secondary antibody (2 µg/ml).

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  Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  Immunohistological analysis using ab110324 at 1μg/ml staining Cyclophilin F in Human cerebellum tissue (Formalin-fixed, Paraffin-embedded).
  
  Note: immunoactivity is most intense in neuronal cell bodies, most notably in the large Purkinje cells.

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  Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  Lanes 1- 2: Merged signal (red and green). Green - ab110324 observed at 23 kDa. Red - Anti-GAPDH antibody[EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) observed at 37 kDa.
  

  ab110324 was shown to react with Cyclophilin F in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line Human PPIF (Cyclophilin F) knockout HEK-293T cell line ab266077 (knockout cell lysate Human PPIF (Cyclophilin F) knockout HEK-293T cell lysate ab257039) was used. Wild-type HEK-293T and PPIF knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab110324 and Anti-GAPDH antibody[EPR16891] - Loading Control (Anti-GAPDH antibody [EPR16891] - Loading Control ab181602) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Mouse IgG H&L (IRDye^®800CW) preadsorbed (Goat anti-Mouse IgG H&L (IRDye® 800CW) preadsorbed ab216772) and Goat Anti-Rabbit IgG H&L (IRDye^®680RD) preadsorbed (Goat Anti-Rabbit IgG H&L (IRDye® 680RD) preadsorbed ab216777) secondary antibodies at 1 in 20000 dilution for 1 hour at room temperature before imaging.

  All lanes: Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1/10000 dilution

  Lane 1: Wild-type HEK-293T cell lysate at 20 µg

  Lane 2: PPIF knockout HEK-293T cell lysate at 20 µg

  Lane 2: Western blot - Human PPIF (Cyclophilin F) knockout HEK-293T cell line (Human PPIF (Cyclophilin F) knockout HEK-293T cell line ab266077)

  Performed under reducing conditions.

  Predicted band size: 22 kDa

  Observed band size: 23 kDa

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  Flow Cytometry - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  Flow cytometric analysis using ab110324 at 1µg/ml staining Cyclophilin F in HL60 cells (blue). Isotype control antibody (red).

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  This image is courtesy of an anonymous customer review

  Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  Blocked with 5% milk for 1 hour at 25°C.

  Incubated with the primary antibody diluted in PBS-T + 5% milk for 16 hours at 4°C.

  All lanes: Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1/1000 dilution

  Lane 1: WT mouse liver mitochondria lysate at 25 µg

  Lane 2: CypD KO mouse liver mitochondria lysate at 25 µg

  Lane 3: WT mouse liver mitochondria lysate at 35 µg

  Lane 4: CypD KO mouse liver mitochondria lysate at 35 µg

  Lane 5: WT mouse liver mitochondria lysate at 50 µg

  Lane 6: CypD KO mouse liver mitochondria lysate at 50 µg

  Secondary

  All lanes: HRP-conjugated goat anti-mouse IgG polyclonal at 1/4000 dilution

  Developed using the ECL technique.

  Performed under reducing conditions.

  Predicted band size: 22 kDa

  Observed band size: 13 kDa, 20 kDa

  Exposure time: 5s

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  Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324)

  All lanes: Western blot - Anti-Cyclophilin F antibody [E11AE12BD4] (ab110324) at 1 µg/mL

  Lane 1: Isolated mitochondria from Human heart at 5 µg

  Lane 2: Isolated mitochondria from Bovine heart at 1 µg

  Lane 3: Isolated mitochondria from Rat heart at 10 µg

  Lane 4: Isolated mitochondria from Mouse heart at 10 µg

  Lane 5: Isolated mitochondria from HepG2 cells at 20 µg

  Predicted band size: 22 kDa